Abstract
Rabies is a neglected tropical disease caused by the rabies virus (RABV) and it is mostly transmitted to humans by dogs. However, knowledge about host factors and their role in RABV infection in dogs is still limited. This is the first study to identify differentially expressed dog brain proteins involved in naturally occurring rabies virus infection using iTRAQ-based proteomics approach. Here, we have reported 19 differentially expressed proteins (DEPs) in RABV infected dog brain compared to controls and some of these proteins are the first time reported. GO annotation revealed their involvement in various molecular functions and also their participation in cellular processes, biological regulation, and metabolic processes. Further, ingenuity pathway analysis (IPA) predicted the major protein network showing the connection of 13 DEPs and their role in some of the canonical pathways. IPA analysis also predicted their involvement in neurological disease, organismal injury and abnormalities, psychological disorders. Proteomics data were successfully validated at gene expression levels by qRT-PCR for five DEPs including two down-regulated proteins namely DDX3X and SNCB and three up-regulated proteins namely KCTD12, DLA-64 and IgSF. This study provides the list of molecules involved in RABV infection and gives some insight into the molecular pathophysiology involved in this infection. This also suggests that DE proteins like DDX3X may have importance in RABV replication similar to other viruses. However, further experiments are required to confirm the role of these proteins in the molecular mechanism of RABV infection.
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Supplementary Figure 2: Representative results of various tests for rabies virus infection A. the presence of Negri bodies with Seller’s staining method, B. Direct Fluorescent Antibody Test (dFAT) showing fine dusty particles emitting bright apple green color fluorescence were considered positive for the presence of rabies virus antigen, C. The amplification of 762 bp region of nucleoprotein (N) gene and D. 406 bp region of glycoprotein, (G) gene of rabies virus (tif 1401 kb)
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Supplementary Table 1: List of down-regulated and up-regulated dog brain proteins expressed in RABV infection along with their mass spectrometry details (xlsx 15 kb)
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Supplementary Table 2: List of all identified dog brain proteins along with their mass spectrometry details (xlsx 76 kb)
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Behera, S., Pharande, R.R., Reddy, R.R. et al. Quantitative proteomics leads to identify dog brain proteins involved in rabies virus infection: implication in understanding viral pathophysiology. J Proteins Proteom 11, 241–257 (2020). https://doi.org/10.1007/s42485-020-00051-w
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DOI: https://doi.org/10.1007/s42485-020-00051-w